Animal protein-free media for cultivation of cells
View Patent ↗The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.
1. A method for expressing a target protein, comprising the steps of:
a) providing a culture of cells into which a nucleic acid comprising a sequence encoding the target protein has been introduced;
b) culturing the cells in an animal protein-free cell culture medium comprising putrescine and at least one protein hydrolysate, wherein the hydrolysate is derived from a plant or yeast, and wherein the putrescine is present in the animal protein-free cell culture medium in a concentration ranging from about 0.5 to about 10 mg/L.
2. The method of claim 1 , wherein the putrescine is present in the animal protein-free cell culture medium in a concentration ranging from about 2 to about 8 mg/L.
3. The method of claim 1 , wherein the putrescine is present in the animal protein-free cell culture medium in a concentration ranging from about 2 to about 5 mg/L.
4. The method of claim 1 , wherein the putrescine is from a different source than the protein hydrolysate.
5. The method of claim 1 , wherein the target protein is selected from the group consisting of a biologically active part of a blood coagulation factor, a biologically active part of a protein involved in the production of red blood cells and angiogenesis, and a monoclonal antibody.
6. The method of claim 5 , wherein the target protein is Factor VIII or erythropoietin.
7. The method of claim 6 , wherein the target protein is Factor VIII.
8. The method of claim 1 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cell, bacterial cell, and yeast cells.
9. The method of claim 8 , wherein the mammalian cells are CHO or BHK cells.
10. The method of claim 1 , wherein the cell is a CHO cell and the target protein is Factor VIII, or the cell is a BHK cell and the target protein is erythropoietin, or the cell is an Epstein Barr virus transformed immortalized human B cell and the target protein is a human antibody.
11. The method of claim 10 , wherein the cell is a CHO cell and the target protein is Factor VIII.
12. The method of claim 1 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion-cultivation, and chemostat-cultivation.
13. The method of claim 1 , wherein the animal protein-free cell culture medium further comprises cadaverine, spermidine, spermine, agmatine, or ornithine, or a combination thereof.
14. The method of claim 1 , wherein the protein hydrolysate is derived from cereal or soy.
15. The method of claim 1 , wherein the protein hydrolysate is soy hydrolysate in a concentration ranging from about 0.5 to 30 mg/L.
16. The method of claim 1 , wherein the protein hydrolysate is present in the animal protein-free cell culture medium in a total concentration ranging from about 0.05 (w/v) to about 5% (w/v) for all protein hydrolysates.
17. The method of claim 1 , wherein the cells express the target protein.
18. The method of claim 1 , further comprising the step of isolating the target protein from the animal protein-free cell culture medium.
19. The method of claim 18 , wherein the target protein is Factor VIII.